JP5840703B2 - Water-soluble composition comprising curcumin having enhanced bioavailability and process for its preparation - Google Patents

Description

The present invention relates to water-soluble compositions having enhanced bioavailability and processes for their preparation. More particularly, the present invention relates to a novel water soluble composition containing curcumin with at least one antioxidant, a hydrophilic carrier, and fat and having enhanced bioavailability.

  The compositions of the present invention are useful for the treatment of depression by alleviating the symptoms of depression in humans. The novel water-soluble compositions of curcumin of the present invention with enhanced bioavailability are particularly useful for formulation into oral delivery forms such as dry blends, tablets, capsules and the like.

BACKGROUND OF THE INVENTION Curcumin [1,7-bis (4-hydroxy-3-methoxyphenyl) -1,6-heptadiene-3,5-dione] is a hydrophobic antioxidant that is a powerful antioxidant derived from the spice turmeric. Polyphenol derivatives. Commercially available curcumin contains approximately 77% diferroylmethane, 17% demethoxycurcumin, and 6% bisdemethoxycurcumin. Curcumin is the main active ingredient of turmeric (Curcuma Longa), which has been used since ancient times. Turmeric (turmeric) is a well-known and unique herbal medicine. Turmeric is known for a variety of biological and pharmacological activities, including anti-inflammatory properties, antioxidant properties, anti-proliferative properties, antibacterial properties, anti-carcinogenic properties, and anti-angiogenic properties.

  Major depression, a debilitating mental disorder, is now considered one of the most prevalent human diseases. Major depression can be caused by a number of reasons, including social, occupational, financial, and interpersonal difficulties. People with depression usually present with a state of sad mood and aversion to everyday activities that generally affect a person's thoughts, behaviors, emotions, and physical health.

  Various antidepressants have been prescribed to alleviate the symptoms of depression. Currently used drugs are mainly monoamine oxidase inhibitors (MAOI), tricyclic antidepressants (TCA), tetracyclic antidepressants (TeCA), selective serotonin reuptake inhibitors (SSRI), and Contains norepinephrine / dopamine reuptake inhibitor (NDRI). Some of the blockbuster drugs that are available on the market and that are indicated for the treatment of depression are sold under trademarks such as Prozac, Norpramin, Effexor, Serzone, Remeron, Desyrel, Zolsoft, paxil, Pamelor, Aventyl, Surmontil ing. Nutritional supplement products such as St Johns Wort are also widely used for depression.

  However, the therapeutic benefits of the above drugs are often accompanied by undesirable side effects and the exact mechanism of action is not well understood. Many associated side effects include nausea, insomnia, anxiety, restlessness, decreased sexual impulses, dizziness, weight gain or loss, tremor, sweating, sleepiness, fatigue, thirst, diarrhea, constipation, headache, and the like. Because of these side effects and because some patients are unable to respond to existing drugs, the need for safer and effective drugs for the treatment of major depression is emphasized.

  Therefore, it would be useful to identify traditional herbal-derived or herbal-derived compositions that would specifically address safety and efficacy issues, thereby alleviating depressive symptoms Conceivable. Curcumin is one such molecule that has shown promising efficacy in various animal models of major depression. The mechanism of curcumin's antidepressant effect is not fully understood but is hypothesized to act through inhibiting the monoamine oxidase enzyme and regulating the release of serotonin and dopamine. In addition, evidence shows that curcumin enhances neurogenesis, particularly in the frontal cortex and hippocampal regions of the brain. (SK Kulkarni et al. Potentials of Curcumin as an Antidepressant; Scientific World Journal 2009 Nov 1; 9: 1233-41 (Non-Patent Document 1)).

  Another study confirmed the antidepressant effect of curcumin in the forced swimming test and suggested that the antidepressant effect could be mediated by actions in the central monoaminergic neurotransmitter system. Curcumin doses of 1.25, 2.5, 5, and 10 mg / kg PO were used in the forced swimming test in rats, and long-term treatment with 14 days of curcumin has been shown to reduce immobility time in the forced swimming test (Ying Xu et al. Antidepressant effects of curcumin in the forced swim test and olfactory bulbectomy models of depression in rats. Pharmacology Biochemistry and behavior. 82 (1) 2005. pp. 200-206 (non-patent document 2)).

  Oral administration (140-560 mg / kg P.O.) of aqueous turmeric extract for 14 days shows a decrease in immobility in the tail suspension test and forced swimming test. The results suggest that the turmeric extract had a specific antidepressant effect in vivo (ZF Yu, LD Kong and Y Chen. Antidepressant activity of aqueous extracts of Curcuma longa in mice. Journal of ethnopharmacology. 83 ( 1-2) 2002.pp.161-165 (Non-Patent Document 3)).

  Studies have shown that stress-induced damage to hippocampal neurons can contribute to the pathophysiology of depression. Curcumin administration (10 and 20 mg / kg, PO) increases hippocampal neurogenesis in chronically stressed rats and exhibits similar activity to imipramine treatment, a classic antidepressant (Ying Xu et al. Curcumin reverses impaired hippocampal neurogenesis and increases serotonin receptor 1A mRNA and brain-derived neurotrophic factor expression in chronically stressed rats. Brainsearch. 1162 (2007) 9-18 (non-patent document 4)).

  Other studies have investigated the involvement of the monoaminergic system in curcumin's antidepressant activity, and the effect of piperine, a bioavailability enhancer, on the bioavailability and biological effects of curcumin. This study shows that curcumin administration inhibited the immobility period, increased serotonin (5-HT) and dopamine levels, and inhibited the monoamine oxidase enzyme in mice. Simultaneous administration of piperine and curcumin resulted in enhanced pharmacological, biochemical, and neurochemical activity (SK Kulkarni et al. Antidepressant activity of curcumin: involvement of serotonin and dopamine system. Psychopharmacology (2008) 201: 435-442 (Non-Patent Document 5)).

  It has recently been suggested that the AC-cAMP second messenger pathway plays an important role in depression. Thus, compounds that control the signal pathway may have potential as antidepressants. The effects of chronic unpredictable mild stress (CUMS) and curcumin on behavioral / serotonergic receptor-coupled AC-cAMP signaling pathways have been studied in rats. Curcumin enhances AC activity and c-AMP levels in platelets and various brain regions and binds AC 2, AC 8, and cAMP response elements, which are AC subtypes in the hippocampus, cortex, and hypothalamus of CUMS rats Protein (CREB) mRNA expression was upregulated. The strong antidepressant properties of curcumin may be attributed to improvements in the AC-cAMP pathway and CREB by inhibiting central 5-HT (1A / 1B / 7) receptors in CUMS rats (YC Li et al. Prog Neuropsychopharmacol Biol Psychiatry. 2009 Apr 30; 33 (3): 435-49 (non-patent document 6)). al. Antidepressant-like effects of curcumin on serotonergic receptor-coupled AC-cAMP pathway in chronic unpredictable mild stress

  In addition to the above, there are numerous clinical studies that discuss the effectiveness of curcumin in humans. Despite clinical data showing strong intrinsic activity suggesting the potential of curcumin to be used as a therapeutic agent, clinics for treatment for a number of illnesses, including major depression, due to its poor gastrointestinal absorption The use of curcumin in is limited.

  The reason for the reduced bioavailability of any agent in the body is due to low intrinsic activity, poor absorption, high metabolic rate, metabolite inactivity, and / or rapid removal and clearance from the body there's a possibility that. Studies on curcumin regarding curcumin absorption, distribution, metabolism, and excretion reveal that curcumin poor absorption and rapid metabolism significantly reduce its bioavailability.

  WO 2007/103435 has an enhanced bioavailability, curcuminoid, antioxidant useful for treating Alzheimer's disease and other age-related disorders Disclosed is a curcuminoid formulation comprising a glucuronidation inhibitor, and a water-soluble pharmaceutically acceptable inhibitor.

  International Publication No. 2008/113177 (Patent Document 2) discloses various compounds and compositions including polyunsaturated fatty acid monoglycerides and their derivatives. These compounds have been shown to be useful for enhancing the solubility of various active agents and enhancing their bioavailability.

  Indian patent application No. 1776 / DEL / 2008 has improved drug availability, improved bioavailability with reasonable physical and chemical stability as a self-nanoemulsifying composition Disclosed are pharmaceutical compositions of curcuminoids.

  Another Indian Patent Application No. 1827 / DEL / 2008 provides curcumin nanoparticles, curcumin bound to chitosan nanoparticles, and methods of making them. The bioavailability of curcumin in these formulations was shown to improve more than 10 times.

  Curcumin in serum by continuous metabolism in the liver resulting in some limitations of these compositions, such as excessive damage to curcumin by cytochrome in the small intestine, less potent curcumin glucuronide and curcumin sulfate formation There are reduced levels, as well as difficulties passing through the blood brain barrier. For these reasons, none of the current technologies have been commercially successful as a treatment for mental depression or as a mood booster.

  Currently used antidepressants have many limitations. In addition to being a prescription drug that requires continuous medical management during treatment, they have serious side effects. Selective serotonin reuptake inhibitors have side effects such as nausea, diarrhea, agitation, loss of sexual impulse and the like. Tricyclic inhibitors have side effects such as dry mouth, visual acuity, dizziness, tremor. Monoamino oxidase inhibitors have side effects such as hepatitis, heart attack, stroke, stroke and the like. Curcumin, derived from dietary sources, does not have such serious side effects.

International Publication No. 2007/103435 International Publication No. 2008/113177 Indian Patent Application No. 1776 / DEL / 2008 Indian Patent Application No. 1827 / DEL / 2008

SK Kulkarni et al. Potentials of Curcumin as an Antidepressant; Scientific World Journal 2009 Nov 1; 9: 1233-41 Ying Xu et al. Antidepressant effects of curcumin in the forced swim test and olfactory bulbectomy models of depression in rats.Pharmacology Biochemistry and behavior. 82 (1) 2005. pp.200-206 ZF Yu, LD Kong and Y Chen. Antidepressant activity of aqueous extracts of Curcuma longa in mice.Journal of ethnopharmacology. 83 (1-2) 2002.pp.161-165 Ying Xu et al. Curcumin reverses impaired hippocampal neurogenesis and increases serotonin receptor 1A mRNA and brain-derived neurotrophic factor expression in chronically stressed rats. Brainsearch. 1162 (2007) 9-18 SK Kulkarni et al. Antidepressant activity of curcumin: involvement of serotonin and dopamine system.Psychopharmacology (2008) 201: 435-442 YC Li et al. Antidepressant-like effects of curcumin on serotonergic receptor-coupled AC-cAMP pathway in chronic unpredictable mild stress of rats.Prog Neuropsychopharmacol Biol Psychiatry. 2009 Apr 30; 33 (3): 435-49

OBJECTIVES OF THE INVENTION Accordingly, the main objective of the present invention is to provide novel water soluble compositions with enhanced bioavailability that are useful as antidepressants in humans.

  Another object of the present invention is to provide a novel water-soluble composition with enhanced bioavailability containing curcumin that can be utilized in an orally administrable form.

  Yet another object of the invention is to contain curcumin and have enhanced bioavailability useful as an antidepressant in humans, which is safer for human consumption without any side effects. It is to provide a novel water-soluble composition.

  Yet another object of the present invention is a novel water-soluble composition containing curcumin and having enhanced bioavailability useful as an antidepressant in humans, with better efficacy than conventional antidepressants Is to provide things.

  Yet another object of the present invention is to provide a process for the preparation of novel water soluble compositions containing enhanced bioavailability and containing curcumin that are useful as antidepressants in humans. is there.

  The present invention is based on sustained R & D performed by the inventors due to the fact that curcumin is combined with antioxidants, hydrophilic carriers, and fats to dramatically enhance the bioavailability of curcumin. Developed based on discovery. To date, no such combination has been known that results in enhanced bioavailability useful for alleviating the symptoms of depression.

SUMMARY OF THE INVENTION Accordingly, the present invention is a novel having enhanced bioavailability useful for the treatment of depression comprising a synergistic combination of curcumin, at least one antioxidant, a hydrophilic carrier, and fat. A water soluble composition is provided.

In accordance with another aspect of the present invention, a process is provided for the preparation of a novel water soluble composition with enhanced bioavailability useful for treating depression comprising the following steps:
(I) dissolving curcumin, at least one antioxidant, a hydrophilic carrier, and fat in a solvent to form a homogeneous mass;
(Ii) warming the resulting mass at a temperature in the range of 25 ° C. to 60 ° C. for 4 to 8 hours to obtain a dry wet mass;
(Iii) removing the solvent by evaporation to form a dry mass; and (iv) pulverizing the dry mass to form a fine powder.
[Invention 1001]
A novel water-soluble composition with enhanced bioavailability useful for the treatment of depression comprising curcumin with at least one antioxidant, a hydrophilic carrier, and fat.
[Invention 1002]
The novel water-soluble composition of curcumin of the invention 1001 wherein the amount of curcumin used in the composition ranges from 1% to about 90% by weight of the total composition.
[Invention 1003]
The novel water-soluble composition of curcumin of the present invention 1001 and 1002 wherein the concentration of antioxidant used is in the range of 0.1% to 10% by weight of the total composition.
[Invention 1004]
A novel water-soluble composition of curcumin according to the invention 1001 to 1003, wherein the concentration of antioxidant used is in the range of 0.1% to 10% by weight of the total composition.
[Invention 1005]
The novel water-soluble composition of curcumin according to the invention 1001 to 1004, wherein the concentration of the hydrophilic carrier used is in the range of 1% to 85% by weight of the total composition.
[Invention 1006]
A novel water-soluble composition of curcumin according to the invention 1001 to 1005, wherein the concentration of fat used ranges from 0.1% to 25% by weight of the total composition.
[Invention 1007]
Inventions 1001 to 1006 wherein the antioxidant used is selected from the group consisting of natural tocopherol, ascorbyl palmitate, rosemary extract, epigallocatechin gallate, catechin, ascorbic acid, and / or mixtures thereof New water-soluble composition of curcumin.
[Invention 1008]
A book wherein the hydrophilic carrier used is selected from the group consisting of soluble starch, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, polyethylene glycol, glycerol, sorbitol, mannitol, glucose, sugar, and / or mixtures thereof. A novel water-soluble composition of curcumin according to inventions 1001 to 1007.
[Invention 1009]
The novel water-soluble composition of curcumin according to the invention 1001 to 1008, wherein the fat used is selected from milk fat, medium chain triglycerides, long chain triglycerides, hydrogenated vegetable oils, and / or mixtures thereof.
[Invention 1010]
A method for preparing a novel water-soluble composition with enhanced bioavailability useful for the treatment of depression comprising the following steps:
(I) dissolving curcumin, at least one antioxidant, a hydrophilic carrier, and fat in a solvent to form a homogeneous mass;
(Ii) heating the resulting mass to a temperature in the range of 25 ° C. to 60 ° C. for 4 to 8 hours to obtain a dry wet mass;
(Iii) removing the solvent by evaporation to form a dry mass; and
(Iv) A step of pulverizing the dry mass to form a fine powder.
[Invention 1011]
The method of the present invention 1010 wherein the curcumin used has a purity content in the range of 50-99%.
[Invention 1012]
The present invention 1010 and 1011 wherein the antioxidant used is selected from the group consisting of natural tocopherol, ascorbyl palmitate, rosemary extract, epigallocatechin gallate, catechin, ascorbic acid, and / or mixtures thereof the method of.
[Invention 1013]
A book wherein the hydrophilic carrier used is selected from the group consisting of soluble starch, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, polyethylene glycol, glycerol, sorbitol, mannitol, glucose, sugar, and / or mixtures thereof. Invention 1010-1012 Method.
[Invention 1014]
The method of the present invention 10101-13, wherein the fat used is selected from milk fat, medium chain triglycerides, long chain triglycerides, hydrogenated vegetable oils, and / or mixtures thereof.
[Invention 1015]
The process of the invention 1010 to 1014, wherein the solvent used is selected from isopropyl alcohol, acetone, methanol, alcohol, and / or mixtures thereof.

DETAILED DESCRIPTION The curcumin used in step (i) can be commercially available with analytical results in the range of 85-96%. Curcumin can also be a turmeric extract rich in curcumin. The amount of curcumin added may be sufficient to produce water-soluble curcumin with an analytical result of 1-55% curcumin.

  The antioxidant used in step (i) may be selected from natural tocopherol, ascorbyl palmitate, rosemary extract, epigallocatechin gallate, catechin, ascorbic acid, and mixtures thereof. The amount of antioxidant used can range from 1 to 10%.

  The hydrophilic carrier used in step (i) is selected from soluble starch, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, polyethylene glycol 200-20000, glycerol, sorbitol, mannitol, glucose, sugar, and mixtures thereof. obtain. The amount of hydrophilic carrier added can range from 10 to 90%.

  The fat used in step (i) may be selected from milk fat, medium chain triglycerides, long chain triglycerides, hydrogenated vegetable oils, and mixtures thereof. The amount of fat used can range from 1 to 25%.

  The solvent used for dissolution in step (i) may be selected from isopropyl alcohol, acetone, methanol, alcohol, and mixtures thereof. The temperature maintained to obtain a homogeneous mass can range from ambient temperature to 70 degrees Celsius; preferably 25 ° C to 60 ° C.

  Removal of the solvent in step (ii) can be done in a vacuum distillation or evaporation technique or by a spray drying technique. The resulting dry mass is pulverized by using a mortar and pestle, mixer-grinder, multi-mill, ball mill, jet mill or the like.

  The beneficial effects of curcumin are well known. However, there are many problems associated with curcumin bioavailability when delivered in oral form. Most of the ingested curcumin is unmetabolized and excreted through the stool, and a small part of the absorbed curcumin is converted into other metabolites and excreted. Curcumin does not easily penetrate the gastrointestinal tract and is under the influence of the liver and other intestinal enzymes. With these enzymes, curcumin in the body is rapidly metabolized, thus reducing its bioavailability in the body. A small amount of curcumin that enters the bloodstream is rapidly metabolized by the liver and kidneys. As such, curcumin is highly lipophilic (and therefore easily crosses the blood brain barrier), but only very small amounts of orally administered curcumin are recorded in serum and brain tissue.

  Cytochrome P450 is a phase I metabolic isozyme required for the metabolism of toxic chemicals such as heterocyclic amines that induce carcinogenic DNA adduct formation. Curcumin has been found to enter the gastrointestinal tract and inhibit cytochrome P450 when ingested throughout the body. As noted above, there has been research done to increase the bioavailability of curcumin when used with piperine. Piperine is a bioenhancer that inhibits cytochrome P450, thereby preventing curcumin metabolism in the body. The compositions of the present invention have been found to enhance bioavailability without the presence of any additional bioenhancers.

  The water-soluble composition of curcumin of the present invention includes an antioxidant, a hydrophilic carrier, and fat. Antioxidants associated with curcumin inhibit cytochrome P450. On the other hand, liver microsomal enzymes or other intestinal enzymes attack only aqueous compounds, so the presence of a fat coating on the composition prevents the composition from attack by these enzymes. Thus, antioxidants and fats play a vital role in enhancing the bioavailability of curcumin.

  Details of the invention are set forth in the following examples, which are provided to illustrate the invention and therefore should not be construed to limit the scope of the invention.

Example 1
18 g curcumin (95%), 0.75 g ascorbyl palmitate, 1.1 g green tea extract containing 0.55 g EGCG, 0.8 G natural tocopherol, 10 g HPMC, 265 g polyvinylpyrrolidone (K 30), and 30 g of medium chain triglyceride was suspended in 600 g of isopropyl alcohol to obtain a homogeneous mass. The resulting homogeneous mass is then heated to 70 degrees Celsius to obtain a dry wet mass that is then subjected to distillation at 600 mm Hg under reduced pressure to remove isopropyl alcohol and dried. I got a trout. The dried mass was then ground in a mixer-grinder to form a fine yellow water soluble powder containing 5.8% curcumin.

Example 2
72 g curcumin (95%), 8 g natural tocopherol, 6 g ascorbyl palmitate, 18 g hydroxypropyl methylcellulose, 15 g hydrogenated soybean oil, and 200 g mannitol are suspended in 500 g ethyl alcohol and the mixture is Obtained. The mixture was then homogenized and heated at 60 degrees Celsius to obtain a homogenized mass. This homogenized mass was subjected to evaporation under vacuum to remove ethyl alcohol to give 317 g of dry mass. The resulting dry mass was then ground with a pestle in a mortar to give a yellow powder with 20.1% curcumin.

Example 3
5 g green tea extract containing 275 g curcumin (95%), 50% EGCG, 10 g ascorbyl palmitate, 30 g medium chain triglyceride, 20 g hardened soybean oil, and 175 g polyvinylpyrrolidone, 500 g ethyl alcohol Suspended in. The mixture was homogenized and heated at 60 degrees Celsius. The resulting mixture was then subjected to evaporation under vacuum to remove ethyl alcohol, yielding 520 g of dry mass. The resulting dried mass was then ground with a pestle in a mortar to give a yellow powder with 51.3% curcumin.

Example 4
500 g of 274 g curcumin (95%), 5.1 g green tea extract containing 50% EGCG, 10.6 g ascorbyl palmitate, 31 g medium chain triglyceride, 20 g hydrogenated soybean oil, and 175 g polyethylene glycol 6000 Suspended in acetone. The mixture was homogenized and heated at 55 degrees Celsius to obtain a homogenized mass. The resulting mixture was subjected to evaporation of acetone under vacuum to give 523 g of dry mass. The mass was then ground with a pestle in a mortar to give a yellow powder with 50.6% curcumin.

Example 5
Curcumin Antidepressant Activity Test Procedure Animal: Male Laca Mouse Method: Paw Salt Forced Swim Test (also called behavioral despair test)
This is a test used to measure the effect of antidepressants on the behavior of laboratory animals, typically rats or mice. Animals are subjected to two trials that force them to swim in an acrylic glass cylinder that is filled with water and cannot escape from it. The first test lasts 15 minutes. Then, after 24 hours, a second test lasting 5 minutes is performed. In the second test, the time spent by the animal without moving is measured. This immobility time is reduced by antidepressants.

Male laca mice maintained under standard laboratory conditions with free access to food and water, rectangular glass jars (25 x 12) with 15 cm water maintained at 24 ± 1 ° C × 25 cm ³ ) After a while, the mouse gives up trying to escape from the water, calms down and becomes immobile. Measure the duration of immobility. The animal was considered immobile whenever the animal was in a slightly rounded but upright position and passively floating in the water with the nose above the surface of the water. Many antidepressants have been shown to reduce the duration of immobility when administered to mice. The total immobility period during the 6 minute test was recorded with the help of a stopwatch.

Test substances: The following drugs were used:
1． Conventional curcumin powder (CC),
2． Compositions of the present invention prepared by the process illustrated in Examples 1-5. The percentage of curcumin dry nutrient system (hereinafter referred to as DNS) composition used in a given example is 5%, 20% and 50%, as follows: DNS 5% Examples 5-6 as 2502-DNS-5B (C-5); DNS 20% as 2502-DNS-20B (C-20) and DNS 50% as 2502-DNS-50B (C-50) And supplied by the applicant;
3． Fluoxetine; other commercially available antidepressant testing drugs such as venlafaxine, desipramine, and tranylcypromine.

A. Testing for curcumin's antidepressant activity Testing for curcumin's antidepressant activity was performed using conventional curcumin (CC) as well as the newer compositions of the present invention, namely 2502-DNS-5B (C-5); Using 3 doses of 20B (C-20); and 2502-DNS-50B (C-50), 50 mg / kg, 100 mg / kg, and 200 mg / kg, and 10 mg / kg fluoxetine (Flx) I went. Tables 1, 2, and 3 below provide data on the immobility duration and percentage of immobility reduction in animals when administered at doses of 50 mg / kg, 100 mg / kg, and 200 mg / kg, respectively. I will provide a. As shown in the figures accompanying this specification, immobility period data obtained from Tables 1, 2, and 3 are plotted against administered dose in Graphs 1, 2, and 3, respectively.

Test substances: 50 mg / kg curcumin (conventional) and the compositions of the present invention 2502-DNS-5B (C-5); 2502-DNS-20B (C-20) and 2502-DNS-50B (C -50).
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 1)

^* p <0.05 when compared to vehicle-treated controls

Test substance: 100 mg / kg curcumin (conventional) and newer formulations 2502-DNS-5B (C-5); 2502-DNS-20B (C-20) and 2502-DNS-50B (C-50) ).
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 2)

^* p <0.05 when compared to vehicle-treated controls

Test substance: 200 mg / kg curcumin (conventional) and newer formulations 2502-DNS-5B (C-5); 2502-DNS-20B (C-20) and 2502-DNS-50B (C-50) ).
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 3)

^* p <0.05 when compared to vehicle-treated controls

  Curcumin compounds C-5, C-20, and C-50, like conventional curcumin (CC), have the greatest response (reduction of immobility time) in the forced swim test (FST) at 45 and 60 minutes. This is shown by the above study. Therefore, 45 minutes is employed as the time interval in further studies to investigate the interaction of these drugs with several types of antidepressants working through various mechanisms of action. All of these formulations at 50 mg / kg showed a uniform immobility period similar to C-C. C-20 and C-50 were more effective at 100 and 200 mg / kg compared to C-5 and C-C. At this dose, the immobility period was similar to that produced by fluoxetine (10 mg / kg).

Combination studies with various antidepressants Curcumin combination studies were conducted with various commercially available antidepressants. Comparison data obtained from these studies are summarized in Tables 4-7. Table 4, Table 5, Table 6, and Table 7 show fluoxetine (5 mg / kg, ip), venlafaxine (2 mg / kg, ip), desipramine (5 mg / kg, ip), and tranylsi, respectively. Comparative data is provided for the duration of immobilization of animals and the percentage decrease in immobility of curcumin (administered at a dose of 50 mg / kg) when administered with promin (5 mg / kg, ip). As shown in the drawings attached to this specification, immobility period data obtained from Tables 4, 5, 6, and 7 are shown in graphs 4, 5, 6, and 7, respectively, for doses administered. And plot.

Combination with less than effective fluoxetine (Table 4)
Test substances: 2502-DNS-5B (C-5); 2502-DNS-20B (C-20) and 2502-DNS-, 50 mg / kg of newer formulations with fluoxetine (5 mg / kg, ip) 50B (C-50), as well as conventional curcumin (CC).
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 4)

^* p <0.05 when compared to vehicle-treated controls

Combination studies with less effective venlafaxine (Table 5)
Test substances: C-5 (2502-DNS-5B), C-20 (2502-DNS-20B), 50 mg / kg newer formulations with venlafaxine (2 mg / kg, ip); and C -50 (2502-DNS-50B), as well as conventional curcumin.
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 5)

^* p <0.05 when compared to vehicle-treated controls

Combination studies with less effective doses of desipramine (Table 6)
Test substance: 2502-DNS-5B (C-5); 2502-DNS-20B (C-20) and 2502-DNS-, 50 mg / kg newer formulations with desipramine (5 mg / kg, ip) 50B (C-50), as well as conventional curcumin (CC).
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 6)

^* p <0.05 when compared to vehicle-treated controls

Combination study with less than effective dose of tranylcypromine (Table 7)
Test substances: newer formulations of 502-mg / kg with tranylcypromine (5 mg / kg, ip) 2502-DNS-5B (C-5); 2502-DNS-20B (C-20) and 2502- DNS-50B (C-50), as well as conventional curcumin (CC).
Animal: Male Laca mouse.
Test procedure: Paw salt forced swimming test. A rectangular glass bottle (25 x 12 x 25 cm ³ ) with 15 cm of water maintained at 24 ± 1 ° C.

^*p＜0.05、媒質で処置した対照と比較した際 (Table 7)

^* p <0.05 when compared to vehicle-treated controls

Conclusion For combination studies with various antidepressants, 50 mg / kg was adopted as an effective dose. With less than an effective amount of fluoxetine (5 mg / kg, a selective serotonin reuptake inhibitor), all of the 50 mg / kg curcumin formulations of the present invention promoted a reduction in immobility time in FST. With sub-effective amounts of venlafaxine (2 mg / kg, a double reuptake inhibitor of serotonin and norepinephrine), C-20 and C-50 are more effective and are similar to those observed in CC The percentage of inhibition of immobility time was 10% higher for C-20 and 20% higher for C-50 compared to CC. In this study, C-5 did not show any enhancement with venlafaxine. With less than an effective amount of desipramine (5 mg / kg, a tricyclic antidepressant), all three newer formulations of curcumin, namely C-5, C-20, and C-50, were enhanced It showed antidepressant activity. Less than effective amounts of CC failed to show any enhancement with desipramine. Both newer curcumin formulations (C-5, C-20, C-50) or CC show enhanced effects with sub-effective amounts of tranylcypromine (5 mg / kg, nonspecific MAO inhibitor) There wasn't.

B. HPLC data of compounds
To study changes in biological amine levels using HPLC, all of the newer curcumin formulations at 100 mg / kg, namely C-5, C-20, C-50, and conventional curcumin CC The animals were injected and sacrificed 45 minutes later.

Test substances: fluoxetine, standard conventional curcumin (100 mg / kg), newer curcumin formulations (100 mg / kg each).
Animal: Male Laca mouse.
Test Procedure: Animals were sacrificed 45 minutes after drug administration as well as behavioral observation time.
HPLC-ECD detector: mobile phase-phosphate buffer: acetonitrile (87:13), pH 4.5
Column-ODS-3 C-18 column (250 x 4.6 mm ID; 5 μm particle size)

(Table 8)

(Table 9)

(Table 10)

(Table 11)

Conclusion
C-5 showed a significant increase (20.99%) in dopamine levels compared to controls. The increase in dopamine levels compared to CC is 71.13, 51.73, and 33.02 for C-5, C-20, and C-50, respectively. C-5 and C-20 showed a significant increase in 5-HT levels (51.63 and 59.19) compared to the control. The increase in 5-HT levels compared to CC is 41.98, 49.05, and 31.13 for C-5, C-20, and C-50, respectively. C-5 and C-20 showed a significant increase (63.17 and 41.84) in norepinephrine levels compared to controls. The increase in norepinephrine levels was similar to that observed in the CC treated group.

Example 6
Comparative bioavailability studies Bioavailability studies were conducted at the request of the applicant at Amala Cancer Institute, Thrissur, Kerala. Twelve healthy subjects were recruited for research purposes. All volunteers were asked to refrain from curcumin-rich food during the course of the study. The study design chosen was a balanced, open-label, two treatment, two period, single dose bioavailability study.

Twelve subjects were recruited for the study and asked to avoid eating food containing abundant turmeric powder or extract for 24 hours prior to each period. A single dose of curcumin capsule (equivalent to 1 gram of curcuminoid) was used for the study. The following supplements were used for research purposes:
Supplement 1-OmniActive Health Technologies Ltd., India Curcumin extract powder 95% (Curcumin extract powder 95%), ie containing CC (500 mg capsule equivalent to 1 gram of curcuminoid), curcumin extract powder 95% 500 A single dose of 2 mg capsules was administered.
Supplement 2-OmniActive Health Technologies Ltd., India's Curcumin Ultrasol Dry Nutrient System 50% (Curcumin Ultrasol Dry Nutrient System 50%), ie containing C-50 (250 mg capsules equivalent to 1 gram of curcuminoid) Curcumin Ultrasol dry nutrition system 50% 250 mg administered as 4 capsules as a single dose.

  Each supplement was administered with 240 ml of water in each period.

methodology:
The primary objective of this study was the bioavailability of curcumin after administration of a single dose of 95% of two curcumin extract powders (500 mg capsules equivalent to 1 gram of curcuminoid) in healthy human subjects Was compared to 4 curcumin Ultrasol dry nutrition systems 50%, ie C-50 (250 mg capsules equivalent to 1 gram of curcuminoid). The second objective of the study was to monitor the safety and tolerability of a single dose of 1 g curcuminoid when administered in 12 healthy human subjects.

  The study design is an open-label, two-treatment, two-period, single-dose bioavailability study. In this study, 12 subjects were recruited and all of them completed the study. Subjects were asked to avoid eating food containing abundant turmeric powder or extract for 24 hours prior to each period. Single dose of curcumin capsule equivalent to 1 gram of curcuminoid: Supplement 1 or Supplement 2 was administered with 240 ml of water each period. There was a washout period of at least 2 weeks between each dose.

  Blood was collected from each subject immediately prior to administration and at 1, 2, 4, 6, 8, and 24 hours after administration. The total blood loss for each subject was approximately 70 ml and there were a total of 14 blood samples of 5 ml at each time point throughout the study. Blood samples taken at various time intervals are centrifuged and curcumin in plasma is measured by HPLC technique.

  The primary efficacy variables measured in this study were Cmax, AUC0-t, and AUC0-∞, and secondary efficacy variables were Tmax, T1 / 2, and Kel as follows: . Statistical evaluation was performed with Winnonlin Software 5.0.1 version. Result variables: From the plasma sample analysis, time vs concentration was plotted, from which Cmax (the maximum concentration of curcumin in the blood) was calculated. AUC for 24 hours was calculated from the obtained peak. Tmax and T1 / 2 were also recorded and compared for the two products.

Analysis method:
Preparation of standard stock solution (157489.2 ng / ml) Standard name: Turmeric standardized extract with standard purity of 96.03% (from Kancor, Angamaly, Kerala).
16.4 mg of turmeric standardized extract was weighed into a 100 ml volumetric flask. Dissolve in methanol and make up to volume.

Preparation of standard solution A (15748.92 ng / ml):
10.0 mL of the standard stock solution was taken and diluted to 100 mL with methanol.

Preparation of standard solution B (787.446 ng / ml):
1.0 mL of standard solution A was taken and diluted to 20 mL with methanol.

Preparation of final standard solution C (393.723 ng / ml):
5.0 mL of standard solution B was taken and diluted to 10 mL with methanol.

Preparation of standard solution D (3149.784 ng / ml):
5.0 mL of Solution A was taken and diluted to 25 mL with methanol.

Preparation of spike standard solution E (1574.892 ng / ml):
5.0 mL of Solution D was taken and diluted to 10 mL with methanol.

Preparation of blank (spike with standard solution E):
Each blank plasma sample was allowed to reach room temperature. 1.0 ml of blank plasma sample and 100 μl of spike standard solution E were mixed in a clean glass test tube. The spiked blank solution was then extracted using the same procedure as the sample.

Preparation of standard (spike with standard solution E):
Pipette 900 μl of blank plasma and 100 μl of standard solution D and transfer into a clean glass test tube. To this, 100 μl of spike standard solution E was added and mixed. This spiked standard was then extracted using the same procedure as the sample.

Preparation of sample (spike with standard solution E):
The frozen plasma sample was allowed to reach room temperature. 1000 μl of plasma sample was pipetted and spiked with 100 μl of spike standard solution E. This solution was vortexed for 1 minute and 3.0 ml of ethyl acetate (HPLC grade) was added to the above solution. The solution was again vortexed for 1 minute with the help of a cyclomixer and allowed to settle at room temperature.

  1.5 ml of the upper ethyl acetate layer was removed with a pipette and evaporated to dryness using vacuum. The dried sample was dissolved in 600 μl methanol (HPLC grade) using a vortex mixer. This solution was filtered through 0.2 micron membrane filter paper. 100 μl of these filtered solutions were injected into HPLC (Waters Alliance System with 2996 PDA detector in no gradient mode). The column used was LiChrospher 100 RP-18 (250 × 4.6 mm 5 μM particle size), methanol was used as the mobile phase, and the detection wavelength was 420 nm. In order to identify and quantize curcuminoids in plasma, chromatographic peaks were compared to standard solutions.

result:
The results from the studies conducted as described above are summarized in the table. Tables 12-17 below show the data obtained.

ND‐血漿中で検出不能 (Table 12) Table of individual concentrations for 95% curcumin extract powder (ng / mL)

ND-not detectable in plasma

Table 13: Table of average concentrations for curcumin extracted powder (minimum 95% curcuminoid)

ND‐検出不能 (Table 14) PK parameters for curcumin extracted powder (minimum 95% curcuminoid)

ND-undetectable

ND‐検出不能 Table 15: Individual concentration table (ng / mL) for the UltraSol nutrition system (50% curcuminoids)

ND-undetectable

Table 16: Average concentration table for UltraSol nutrition system (50% curcuminoids)

ND‐検出不能 Table 17: PK parameters for UltraSol nutrition system (50% curcuminoids)

ND-undetectable

Conclusion Prior art shows that curcumin is not well absorbed upon oral administration. With oral administration of 3.6 g of curcumin, approximately 11 ng / ml can be measured in plasma. Special efforts / techniques are required to detect curcumin in plasma at lower doses. However, in this study, 1 g of curcumin was orally administered to all subjects over two periods for both supplements. As mentioned above, this 1 gram dose was very low to detect curcumin in plasma samples and the plasma sample was spiked externally with 25 ng curcumin to facilitate its measurement in HPLC. After quantification of curcumin in the plasma, the area of curcumin spiked from the outside of 25 ng was subtracted from the total area of the chromatogram to reach the curcumin concentration.

  The results of bioanalysis showed that curcumin was absent in most of the blood sampling time points in the curcumin supplemented group. In the case of UltraSol DNS curcumin supplementation, curcumin at almost all sampling points was measurable and average serum concentrations up to 21 ng / ml were measured. Curcumin extract powder 95% (actual curcuminoid content is 95.2%) has 1.98 times higher curcuminoid content compared to UltraSol DNS Curcumin 50% Powder (actual curcuminoid content is 48.17%) . In both curcumin supplements, the curcuminoid content was equivalent, ie 1 gram.

  As shown in Graph 8, the average AUC for Curcumin Ultrasol Nutrient System 50% is 541.93 ng / ml * h and for Curcumin Extract Powder 95% is 80.43 ng / ml * h. Curcumin Ultrasol nutrition system 50% AUC shows a 6.74 fold increase over curcumin extract powder 95%, and Ultrasol nutrition system has higher bioavailability (6.74 fold) compared to curcumin extract powder 95% It shows that.

  Compared to Biocurcumax, 6.93 times higher bioavailability was shown compared to curcumin powder, but this increase is at a dose of 2 grams / d. 2 However, in this study, UltraSol DNS curcumin showed 6.74 times higher bioavailability compared to simple curcumin powder, even at a dose of 1 gram / d, which is half that of Biocurcumax. Therefore, the 50% curcumin dry powder formulation of the Ultrasol nutrition system demonstrates higher absorption compared to 95% curcumin powder at the same dose level.

  The peak median concentration achieved by Ultrasol DNS 50%, ie Tmax is 2 hours, 3 hours for curcumin extract powder 95%, Ultrasol DNS curcumin is acting faster than curcumin powder extract Can be shown.

  Safety: A given dose of 1 gram of curcumin in both 95% powder and Ultrasol DNS 50% form was well tolerated in all subjects and no adverse events were reported throughout the study.

  From the above details, it can be seen that the composition of the present invention is not a mere mixture resulting in a composition having a set of component properties used, but a composition formed by synergistic activity of the components used. Can be observed.

Advantages of the Invention The novel water soluble composition of the present invention comprises:
1． It exhibits enhanced bioavailability typically useful for alleviating depression.
2． It is not toxic.
3． It can be easily formulated in an orally administrable form such as tablets, capsules, mixed powders and the like.
Four. Useful for oral delivery of curcumin at high doses for applications such as antidepressants.

Claims (7)

A method for preparing a water-soluble composition with enhanced bioavailability useful for the treatment of depression comprising the following steps:
(I) dissolving curcumin, antioxidant, hydrophilic carrier, and fat in a solvent to form a homogeneous mass;
(Ii) heating the resulting mass to a temperature in the range of 25 ° C. to 60 ° C. for 4 to 8 hours to obtain a dry wet mass;
(Iii) removing the solvent by evaporation to form a dry mass; and (iv) pulverizing the dry mass to form a water-soluble powder. The method of claim 1 , wherein the curcumin has a purity in the range of 50 wt% to 99 wt%. The method according to claim 1 or 2 , wherein the antioxidant is selected from the group consisting of natural tocopherol, ascorbyl palmitate, rosemary extract, epigallocatechin gallate, catechin, ascorbic acid, and mixtures thereof. The hydrophilic carrier of Claims 1-3 , wherein the hydrophilic carrier is selected from the group consisting of soluble starch, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, polyethylene glycol, glycerol, sorbitol, mannitol, glucose, sugar, and mixtures thereof. The method according to any one of the above. The method according to any one of claims 1 to 4 , wherein the fat is selected from milk fat, medium chain triglycerides, long chain triglycerides, hydrogenated vegetable oils, or mixtures thereof. 6. The method according to any one of claims 1 to 5 , wherein the solvent is selected from isopropyl alcohol, acetone, methanol, alcohol, or mixtures thereof. The method of claim 1 , wherein the bioavailability of curcumin is higher than in a control composition comprising curcumin.

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